PMID- 16835241
OWN - NLM
STAT- MEDLINE
DCOM- 20061031
LR  - 20211203
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 281
IP  - 37
DP  - 2006 Sep 15
TI  - Mapping the GRIF-1 binding domain of the kinesin, KIF5C, substantiates a role for 
      GRIF-1 as an adaptor protein in the anterograde trafficking of cargoes.
PG  - 27216-28
AB  - Gamma-aminobutyric acid, type A (GABAA) receptor interacting factor-1 (GRIF-1) 
      and N-acetylglucosamine transferase interacting protein (OIP) 106 are both 
      members of a newly identified coiled-coil family of proteins. They are 
      kinesin-associated proteins proposed to function as adaptors in the anterograde 
      trafficking of organelles to synapses. Here we have studied in more detail the 
      interaction between the prototypic kinesin heavy chain, KIF5C, kinesin light 
      chain, and GRIF-1. The GRIF-1 binding site of KIF5C was mapped using truncation 
      constructs in yeast two-hybrid interaction assays, co-immunoprecipitations, and 
      co-localization studies following expression in mammalian cells. Using these 
      approaches, it was shown that GRIF-1 and the KIF5C binding domain of GRIF-1, 
      GRIF-1-(124-283), associated with the KIF5C non-motor domain. Refined studies 
      using yeast two-hybrid interactions and co-immunoprecipitations showed that 
      GRIF-1 and GRIF-1-(124-283) associated with the cargo binding region within the 
      KIF5C non-motor domain. Substantiation that the GRIF-1-KIF5C interaction was 
      direct was shown by fluorescence resonance energy transfer analyses using 
      fluorescently tagged GRIF-1 and KIF5C constructs. A significant fluorescence 
      resonance energy transfer value was found between the C-terminal EYFP-tagged 
      KIF5C and ECFP-GRIF-1, the C-terminal EYFP-tagged KIF5C non-motor domain and 
      ECFP-GRIF-1, but not between the N-terminal EYFP-tagged KIF5C nor the EYFP-KIF5C 
      motor domain and ECFP-GRIF-1, thus confirming direct association between the two 
      proteins at the KIF5C C-terminal and GRIF-1 N-terminal regions. 
      Co-immunoprecipitation and confocal imaging strategies further showed that GRIF-1 
      can bind to the tetrameric kinesin light-chain/kinesin heavy-chain complex. These 
      findings support a role for GRIF-1 as a kinesin adaptor molecule requisite for 
      the anterograde delivery of defined cargoes such as mitochondria and/or vesicles 
      incorporating beta2 subunit-containing GABAA receptors, in the brain.
FAU - Smith, Miriam J
AU  - Smith MJ
AD  - Department of Pharmaceutical and Biological Chemistry, School of Pharmacy, 
      University of London, 29/39 Brunswick Square, London WC1N 1AX, United Kingdom.
FAU - Pozo, Karine
AU  - Pozo K
FAU - Brickley, Kieran
AU  - Brickley K
FAU - Stephenson, F Anne
AU  - Stephenson FA
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20060711
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Adaptor Protein Complex 1)
RN  - 0 (Carrier Proteins)
RN  - 0 (Intracellular Signaling Peptides and Proteins)
RN  - 0 (Nerve Tissue Proteins)
RN  - 0 (Receptors, GABA-A)
RN  - 0 (TRAK2 protein, human)
RN  - EC 3.6.1.- (KIF5C protein, human)
RN  - EC 3.6.4.4 (Kinesins)
SB  - IM
MH  - Adaptor Protein Complex 1/chemistry
MH  - Amino Acid Motifs
MH  - Animals
MH  - Brain/metabolism
MH  - COS Cells
MH  - Carrier Proteins/*chemistry/physiology
MH  - Chlorocebus aethiops
MH  - Fluorescence Resonance Energy Transfer
MH  - Humans
MH  - Intracellular Signaling Peptides and Proteins
MH  - Kinesins/*chemistry
MH  - Nerve Tissue Proteins/*chemistry/physiology
MH  - Protein Binding
MH  - Protein Conformation
MH  - Receptors, GABA-A/metabolism
MH  - Two-Hybrid System Techniques
EDAT- 2006/07/13 09:00
MHDA- 2006/11/01 09:00
CRDT- 2006/07/13 09:00
PHST- 2006/07/13 09:00 [pubmed]
PHST- 2006/11/01 09:00 [medline]
PHST- 2006/07/13 09:00 [entrez]
AID - S0021-9258(19)34899-9 [pii]
AID - 10.1074/jbc.M600522200 [doi]
PST - ppublish
SO  - J Biol Chem. 2006 Sep 15;281(37):27216-28. doi: 10.1074/jbc.M600522200. Epub 2006 
      Jul 11.